CN118557794B

CN118557794B - Polypeptide sodium hyaluronate composite solution for injection and preparation method thereof

Info

Publication number: CN118557794B
Application number: CN202411036425.8A
Authority: CN
Inventors: 符运会; 金家铸; 盛江伟; 杨燕楠; 王军茹; 朱华青
Original assignee: Hangzhou Keteng Biological Product Co ltd
Current assignee: Hangzhou Keteng Biological Product Co ltd
Priority date: 2024-07-31
Filing date: 2024-07-31
Publication date: 2025-01-07
Anticipated expiration: 2044-07-31
Also published as: CN118557794A

Abstract

The application relates to the technical field of biomedical materials and cosmetic or cosmeceutical preparations, and particularly discloses a polypeptide sodium hyaluronate composite solution for injection and a preparation method thereof. The polypeptide sodium hyaluronate composite solution consists of a solvent and a solute, wherein the solvent is a buffer solution, and the solute mainly comprises the following raw materials of 5-50mg/mL of sodium hyaluronate, 1-10mg/mL of L-carnosine, 0.05-0.15 mg/mL of acetyl hexapeptide-8, 0.05-0.1 mg/mL of acetyl tetrapeptide-5, and 0.03-0.1 mg/mL of tripeptide-1 derivative, wherein the tripeptide-1 derivative is obtained by treating tripeptide-1 with ethyl palmitate and astaxanthin. The polypeptide sodium hyaluronate composite solution shows better anti-wrinkle and skin tightening effects through the synergy among the raw materials, has the effects of moisturizing, moisturizing and resisting oxidation, can improve the plumpness of skin, and enables the skin to be healthier, better in quality and natural.

Description

Polypeptide sodium hyaluronate composite solution for injection and preparation method thereof

Technical Field

The application relates to the technical field of biomedical materials and cosmetic or makeup preparations, in particular to a polypeptide sodium hyaluronate composite solution for injection and a preparation method thereof.

Background

With the development of society and the improvement of living standard, people pay more attention to the problem of skin aging. After birth, the skin tissue of the human is developed and the functions are active. With age, skin metabolism is gradually slow, collagen and elastin synthesis is reduced and lost, and elasticity is gradually lost, and the skin becomes loose and wrinkles appear. In particular, after the age of 35 years, the skin gradually enters the aging phase. Although skin aging is a natural law and cannot be blocked, we can improve the state of skin aging through a scientific method. At this time, in the cosmetic field, wrinkle resistance and skin tightening have become one of the key demands. At present, various anti-wrinkle skin-tightening products are appeared in the market, such as anti-free radical, plant extracts, chinese herbal medicine extracts and the like, however, the anti-free radical is more, and the extraction modes of the plant extracts or the Chinese herbal medicine extracts are different, so that the effects of the products are different, and the anti-wrinkle skin-tightening products still need to be improved. In view of this, how to improve the anti-wrinkle and skin tightening effects has become one of the technical problems to be solved in the art.

Disclosure of Invention

In order to improve the anti-wrinkle and skin tightening effects, the application provides a polypeptide sodium hyaluronate composite solution for injection and a preparation method thereof.

In a first aspect, the application provides a polypeptide sodium hyaluronate composite solution for injection, which adopts the following technical scheme:

The polypeptide sodium hyaluronate composite solution for injection consists of a solvent and a solute, wherein the solvent is a buffer solution, and the solute mainly comprises 5-50mg/mL of sodium hyaluronate, 1-10mg/mL of L-carnosine, 0.05-0.15 mg/mL of acetyl hexapeptide-8, 0.05-0.1 mg/mL of acetyl tetrapeptide-5, 0.03-0.1 mg/mL of tripeptide-1 derivative, and the tripeptide-1 derivative is obtained by treating tripeptide-1 with ethyl palmitate and astaxanthin.

According to the polypeptide sodium hyaluronate composite solution, through the interaction of the raw materials, the elastase inhibition rate is more than 62%, and the type I collagen up-regulation rate is more than 34%, so that the polypeptide sodium hyaluronate composite solution has the characteristics of good effect of reducing skin wrinkles and strong effect of promoting collagen synthesis. And the polypeptide sodium hyaluronate composite solution is applied to improving skin, the wrinkle improvement rate is more than 33%, and the elasticity index improvement rate is more than 97%, so that the polypeptide sodium hyaluronate composite solution shows better anti-wrinkle skin tightening effect.

The polypeptide sodium hyaluronate composite solution has excellent moisturizing performance and oxidation resistance, wherein sodium hyaluronate and L-carnosine, acetyl hexapeptide-8, acetyl tetrapeptide-5 and tripeptide-1 derivatives play a synergistic role, the anti-wrinkle skin tightening effect is improved, the hydration of skin is promoted, the nutrition of dermal cells is enhanced, the fullness of skin is improved, the elasticity and the skin tightening performance are improved, and the polypeptide sodium hyaluronate composite solution is suitable for moisturizing and moisturizing, so that the skin is healthier, better in quality and natural.

Alternatively, the total content of said acetyl hexapeptide-8, acetyl tetrapeptide-5, tripeptide-1 derivatives is >0.15mg/mL.

By adopting the technical scheme, the total content of the acetyl hexapeptide-8, the acetyl tetrapeptide-5 and the tripeptide-1 derivatives is limited, so that the efficacy of the polypeptide sodium hyaluronate composite solution is ensured.

Optionally, the weight average molecular weight of the sodium hyaluronate is 20-200 kilodaltons. Preferably, the weight average molecular weight of the sodium hyaluronate is 50-150 kilodaltons. Still preferably, the sodium hyaluronate has a weight average molecular weight of 120 kilodaltons.

In various embodiments, the weight average molecular weight of sodium hyaluronate is 120 kilodaltons, which may also be set to 20, 50, 80, 100, 130, 150, 180, 200 kilodaltons as desired, but is not limited to the recited values, as other non-recited values within the range are equally applicable.

Optionally, the buffer solution is phosphate buffer solution, the pH value of the phosphate buffer solution is 6.6-7.6, and the osmotic pressure is 270-330 mOsm/L.

By adopting the technical scheme, the buffer solution is adopted to dilute the solute, so that the preparation of the polypeptide sodium hyaluronate composite solution is facilitated, the buffer solution is phosphate buffer solution, the pH value of the buffer solution is 6.6-7.6, the osmotic pressure of the buffer solution is 270-330 mOsm/L, the buffer solution is adapted to the skin, the biocompatibility is increased, the diffusion of the solute in the skin can be promoted, the solute utilization rate is improved, and the safety, stability and efficacy of the polypeptide sodium hyaluronate composite solution are ensured.

Optionally, the phosphate buffer solution is prepared from disodium hydrogen phosphate, sodium dihydrogen phosphate, sodium chloride and water.

In various embodiments, the phosphate buffer has a pH of 6.68 and an osmotic pressure of 300 mOsm/L, which may also be set to pH 6.6, 6.7, 6.8, 6.9, 7, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, as desired, and 270 mOsm/L, 280 mOsm/L, 290 mOsm/L, 300 mOsm/L, 310 mOsm/L, 320 mOsm/L, 330 mOsm/L, as desired, although other values not recited in this range may be equally suitable.

Optionally, the phosphate buffer is prepared by mixing water, disodium hydrogen phosphate, sodium dihydrogen phosphate and sodium chloride, filtering, and collecting filtrate to obtain phosphate buffer.

Alternatively, an aqueous filter membrane is used for filtration, and the pore size of the aqueous filter membrane is 0.1-0.5 μm. Preferably, the pore size of the aqueous filter is 0.2-0.3. Mu.m. Still more preferably, the pore size of the aqueous filter is 0.22. Mu.m.

Optionally, the tripeptide-1 derivative is prepared by mixing an organic solvent and ethyl palmitate, then dropwise adding a tripeptide-1 ethanol diluent, continuously stirring for reaction 17-23 h after the dropwise addition, then dropwise adding an astaxanthin ethanol diluent, continuously stirring for reaction 27-33 h after the dropwise addition, concentrating under reduced pressure, and drying to obtain the tripeptide-1 derivative.

Optionally, the weight ratio of the tripeptide-1 in the ethyl palmitate and tripeptide-1 ethanol diluent to the astaxanthin in the astaxanthin ethanol diluent is (4-6) (5-9) (4-6).

By adopting the technical scheme, firstly, an ester group in ethyl palmitate reacts with an amino group in tripeptide-1 to form an intermediate, grafting of the ethyl palmitate is realized, and the intermediate contains hexadecyl and amino groups. Then, the amino in the intermediate reacts with carbon-carbon double bond in the astaxanthin to obtain tripeptide-1 derivative, so as to realize the grafting of the astaxanthin. The tripeptide-1 is treated by utilizing ethyl palmitate and astaxanthin, and the elastase inhibition rate and the type I collagen up-regulation rate can be improved by utilizing the synergy between the ethyl palmitate and the astaxanthin, so that the tripeptide-1 derivative acts on the dermis layer, the wrinkle depth and the wrinkle area are reduced, the synthesis of collagen and glycosaminoglycan is promoted, the skin is helped to retain moisture, the skin elasticity is recovered, and the anti-wrinkle skin tightening effect is enhanced.

Optionally, the weight ratio of tripeptide-1 and ethanol in the tripeptide-1 ethanol diluent is 1 (3-7), and the weight ratio of astaxanthin and ethanol in the astaxanthin ethanol diluent is 1 (3-7).

By adopting the technical scheme, the content of tripeptide-1 in the tripeptide-1 ethanol diluent and the content of astaxanthin in the astaxanthin ethanol diluent are limited, so that the preparation of the tripeptide-1 ethanol diluent and the astaxanthin ethanol diluent is facilitated.

The tripeptide-1 ethanol diluent is prepared from tripeptide-1 and ethanol. The astaxanthin ethanol diluent is prepared from astaxanthin and ethanol. In various embodiments, the weight ratio of tripeptide-1 to ethanol in the tripeptide-1 ethanol diluent is 1:5, and the weight ratio can be set to be 1:3, 1:4, 1:6 and 1:7 according to the requirement, but the weight ratio is not limited to the listed values, and other non-listed values in the range of the values are equally applicable. In various embodiments, the weight ratio of astaxanthin to ethanol in the astaxanthin ethanol diluent is 1:5, and the weight ratio can be set to be 1:3, 1:4, 1:6 and 1:7 according to the requirement, but the astaxanthin ethanol diluent is not limited to the listed values, and other non-listed values in the range of the values are equally applicable.

Optionally, the weight ratio of the organic solvent to the ethyl palmitate is (30-70) to (4-6).

By adopting the technical scheme, the weight ratio of the organic solvent to the ethyl palmitate is optimized, the limit of the addition amount of the organic solvent is realized, and the sufficient contact and reaction of the ethyl palmitate, the tripeptide-1 and the astaxanthin are facilitated. In one embodiment, the weight ratio of the organic solvent to the ethyl palmitate is 50:4, in another embodiment, the weight ratio of the organic solvent to the ethyl palmitate is 50:5, in another embodiment, the weight ratio of the organic solvent to the ethyl palmitate is 50:6, and the weight ratio of the organic solvent to the ethyl palmitate can be set as 30:4, 30:5, 30:7, 70:4 and 70:5 according to the requirement, but the weight ratio is not limited to the listed values, and other non-listed values in the range of the values are equally applicable.

Optionally, the organic solvent is one or more of ethanol, methanol, propanol and isopropanol.

By adopting the technical scheme, the organic solvent is optimized, so that the selection of the organic solvent is facilitated. In various embodiments, the organic solvent is ethanol, which may be ethanol, methanol, propanol, isopropanol, if desired, but is not limited to the listed choices, and other non-listed choices are equally applicable within the scope of the choices.

Optionally, the dripping time of the tripeptide-1 ethanol diluent is 20-40 min, and the dripping time of the astaxanthin ethanol diluent is 20-40 min.

By adopting the technical scheme, the dripping time of the tripeptide-1 ethanol diluent and the astaxanthin ethanol diluent is limited, so that the contact and the reaction of ethyl palmitate, tripeptide-1 and astaxanthin are facilitated, and the side reaction is reduced.

In a second aspect, the application provides a preparation method of the polypeptide sodium hyaluronate composite solution for injection, which adopts the following technical scheme:

the preparation method of the polypeptide sodium hyaluronate composite solution for injection comprises the following steps:

S1, preparing a buffer solution, filtering and sterilizing to obtain a sterile buffer solution;

S2, mixing sterile buffer solution, sodium hyaluronate, L-carnosine, acetyl hexapeptide-8, acetyl tetrapeptide-5 and tripeptide-1 derivatives in a sterile environment to obtain a polypeptide sodium hyaluronate composite solution.

By adopting the technical scheme, the preparation of the polypeptide sodium hyaluronate composite solution is facilitated.

Optionally, after the polypeptide sodium hyaluronate composite solution is obtained in the step S2, canning the polypeptide sodium hyaluronate composite solution in a sterile environment to obtain a canned product.

Optionally, the canned container used in canning is one of a prefilled syringe, an ampoule bottle and a penicillin bottle.

By adopting the technical scheme, the polypeptide sodium hyaluronate composite solution is canned, so that a canned product is obtained, and the polypeptide sodium hyaluronate composite solution is convenient to store and use.

In summary, the application has at least the following advantages:

1. The polypeptide sodium hyaluronate composite solution provided by the application has the advantages that the synergistic effect is exerted between sodium hyaluronate and L-carnosine, acetyl hexapeptide-8, acetyl tetrapeptide-5 and tripeptide-1 derivatives, skin wrinkles can be effectively reduced, collagen synthesis is promoted, the elastase inhibition rate is more than 62%, the type I collagen up-regulation rate is more than 34%, the wrinkle improvement rate is more than 33%, the elasticity index improvement rate is more than 97%, and a better anti-wrinkle skin tightening effect is shown. The polypeptide sodium hyaluronate composite solution has the effects of moisturizing, resisting oxidation, resisting wrinkle and tightening skin, can promote the hydration of skin, enhance the nutrition to dermal cells, improve the plumpness of the skin, increase the elasticity of the skin and tighten the skin, and ensure that the skin is healthier, better in quality and natural.

2. In the preparation method of the tripeptide-1 derivative, firstly, ethyl palmitate and tripeptide-1 are reacted to realize grafting of the ethyl palmitate to obtain an intermediate, and then, the intermediate and astaxanthin are reacted to realize grafting of the astaxanthin to obtain the tripeptide-1 derivative. The preparation of the tripeptide-1 derivative is facilitated by the reaction and grafting in steps, the quality of the tripeptide-1 is guaranteed, the use effect of the tripeptide-1 is improved, the elastase inhibition rate and the type I collagen up-regulation rate are increased, and the wrinkle improvement rate and the elasticity index improvement rate are improved.

Detailed Description

In order that the application may be more readily understood, the application will be further described in detail with reference to the following examples, which are given by way of illustration only and are not limiting in scope of application. The starting materials or components used in the present application may be prepared by commercial or conventional methods unless specifically indicated.

Preparation example

Preparation example 1

A tripeptide-1 derivative prepared by the following method:

At a rotation speed of 500 r/min, ethyl palmitate of 5g was added to an organic solvent of 50: 50 g, and the mixture was stirred for 5: 5 min. Then 42 g of tripeptide-1 ethanol diluent is added dropwise, and the tripeptide-1 ethanol diluent is added dropwise after 30 min. And after the dripping is finished, stirring the mixture continuously for 20 h. Then, 31.2 g of astaxanthin ethanol diluent is added dropwise, and the astaxanthin ethanol diluent is added dropwise after 30. 30 min. And after the dripping is finished, stirring for 30 h. Concentrating under reduced pressure, and oven drying to obtain tripeptide-1 derivative.

The organic solvent is ethanol, the tripeptide-1 ethanol diluent is prepared from tripeptide-1 and ethanol, the weight ratio of tripeptide-1 to ethanol in the tripeptide-1 ethanol diluent is 1:5, the astaxanthin ethanol diluent is prepared from astaxanthin and ethanol, and the weight ratio of astaxanthin and ethanol in the astaxanthin ethanol diluent is 1:5.

Preparation example 2

A tripeptide-1 derivative prepared by the following method:

Ethyl palmitate of 4 g is added to an organic solvent of 50 g at a rotation speed of 500 r/min, and the mixture is stirred for 5 min. Then 54 g of tripeptide-1 ethanol diluent is added dropwise, and the tripeptide-1 ethanol diluent is added dropwise at 40 min. And after the dripping is finished, continuing stirring treatment 23 h. Then, 24 g of astaxanthin ethanol diluent is dripped, and the astaxanthin ethanol diluent is dripped after 20 min. And after the dripping is finished, stirring the mixture continuously for 27 h. Concentrating under reduced pressure, and oven drying to obtain tripeptide-1 derivative.

Preparation example 3

A tripeptide-1 derivative prepared by the following method:

At a rotation speed of 500 r/min, ethyl palmitate of 6 g is added into an organic solvent of 50 g, and the mixture is stirred for 5 min. Then, 30 g of tripeptide-1 ethanol diluent is added dropwise, and the tripeptide-1 ethanol diluent is added dropwise after 20 min. And after the dripping is finished, stirring for 17 hours. Then 36 g of astaxanthin ethanol diluent is added dropwise, and the astaxanthin ethanol diluent is added dropwise after 40 min. And after the dripping is finished, stirring for 33 h. Concentrating under reduced pressure, and oven drying to obtain tripeptide-1 derivative.

Examples

TABLE 1 raw materials for polypeptide sodium hyaluronate composite solution

Example 1

The raw materials and the raw material proportions of the polypeptide sodium hyaluronate composite solution for injection are shown in table 1.

Wherein, the weight average molecular weight of the sodium hyaluronate is 120 ten thousand daltons, and the tripeptide-1 derivative is prepared by adopting the method of preparation example 1.

The buffer solution is phosphate buffer solution, the pH value of the phosphate buffer solution is 6.68, the osmotic pressure is 300 mOsm/L, and the phosphate buffer solution is prepared from disodium hydrogen phosphate, sodium dihydrogen phosphate, sodium chloride and water.

The phosphate buffer solution is prepared by adding disodium hydrogen phosphate, sodium dihydrogen phosphate and sodium chloride into water at a rotation speed of 300 r/min, and stirring for 10 min to obtain the phosphate buffer solution.

A preparation method of polypeptide sodium hyaluronate composite solution for injection comprises the following steps:

S1, preparing a buffer solution, filtering by adopting a 0.22 mu m water-based filter membrane, taking a filtrate, putting the filtrate into an autoclave, and sterilizing the filtrate at 121 ℃ for 30 min to obtain the sterile buffer solution.

S2, adding sodium hyaluronate into a sterile buffer solution under the sterile environment at the rotating speed of 300 r/min, mixing and treating for 10min, then adding L-carnosine, acetyl hexapeptide-8, acetyl tetrapeptide-5 and tripeptide-1 derivatives, mixing and treating for 20 min to obtain the polypeptide sodium hyaluronate composite solution.

And S3, canning the polypeptide sodium hyaluronate composite solution in a sterile environment to obtain a canned product, wherein a canning container used for canning is a prefilled syringe.

Example 2

The difference between the sodium hyaluronate composite solution for injection and the embodiment 1 is that the raw material ratio of the sodium hyaluronate composite solution for injection is different, and the raw material ratio of the sodium hyaluronate composite solution for injection is shown in table 1.

Example 3

Example 4

The difference between the sodium hyaluronate composite solution for injection and the embodiment 1 is that the source of the tripeptide-1 derivative in the raw material of the sodium hyaluronate composite solution for injection is different, and the tripeptide-1 derivative is prepared by adopting the method of the preparation example 2.

Example 5

The difference between the sodium hyaluronate complex solution for injection and the example 1 is that the source of the tripeptide-1 derivative in the raw material of the sodium hyaluronate complex solution for injection is different, and the tripeptide-1 derivative is prepared by the method of preparation example 3.

Comparative example

Comparative example 1

A polypeptide sodium hyaluronate complex solution for injection is different from example 1 in that tripeptide-1 derivatives are replaced with tripeptide-1 in the raw material of the polypeptide sodium hyaluronate complex solution in equal amounts.

Comparative example 2

A polypeptide sodium hyaluronate composite solution for injection is different from example 1 in that in the raw material of the polypeptide sodium hyaluronate composite solution, the astaxanthin is replaced by the ethyl palmitate in the same amount in the preparation method of the tripeptide-1 derivative.

Comparative example 3

A polypeptide sodium hyaluronate composite solution for injection is different from example 1 in that in the raw material of the polypeptide sodium hyaluronate composite solution, the ethyl palmitate is replaced by the same amount of astaxanthin in the preparation method of the tripeptide-1 derivative.

Comparative example 4

A polypeptide sodium hyaluronate composite solution for injection is different from example 1 in that no acetyl hexapeptide-8 or tripeptide-1 derivative is added to the raw materials of the polypeptide sodium hyaluronate composite solution.

Comparative example 5

A polypeptide sodium hyaluronate complex solution for injection is different from example 1 in that the tripeptide-1 derivative is replaced by an equivalent amount of acetyl hexapeptide-8 in the raw material of the polypeptide sodium hyaluronate complex solution.

Comparative example 6

A polypeptide sodium hyaluronate complex solution for injection is different from example 1 in that the acetyl hexapeptide-8 is replaced by the same amount of tripeptide-1 derivative in the raw material of the polypeptide sodium hyaluronate complex solution.

Performance detection

(1) According to GB/T16886 biological evaluation of medical instruments, the polypeptide sodium hyaluronate complex solutions obtained in examples 1-5 were subjected to skin sensitization test, intradermal stimulation test, and cytotoxicity test, respectively. Through tests, the polypeptide sodium hyaluronate composite solution obtained in the examples 1-5 is less than or equal to 1 level in skin sensitization tests, less than or equal to 1 level in intradermal stimulation tests and less than or equal to 1 level in cytotoxicity tests, and the polypeptide sodium hyaluronate composite solution obtained in the examples 1-5 meets the safety requirement and has good safety.

(2) Elastase is mainly synthesized and secreted by fibroblasts, has high selectivity and specificity, and can degrade elastin in skin, thereby causing skin aging. Thus, the efficacy of the polypeptide sodium hyaluronate complex solution in reducing skin wrinkles was characterized by the inhibition rate of elastase. The polypeptide sodium hyaluronate complex solutions obtained in examples 1 to 5 and comparative examples 1 to 6 were used to measure the elastase inhibition rate, and the measurement results are shown in Table 2.

The elastase inhibitor is prepared by adding 200 μl of polypeptide sodium hyaluronate complex solution and 50 μl of porcine pancreatic elastase solution into 96-well plate at 25deg.C, and incubating for 15 min. Then 50. Mu.L of N-succinyl-L-alanyl-L-alanine solution was added and incubated 15 min. At wavelength 410 nm, absorbance was measured using an enzyme-labeled instrument and elastase inhibition was calculated.

The enzyme activity concentration of the porcine pancreatic elastase solution is 0.171U/mL, and the mass concentration of the N-succinyl-L-alanyl-L-alanine in the N-succinyl-L-alanyl-L-alanine solution is 90%.

(3) The type I collagen is one of main components of the extracellular matrix of dermis, and is synthesized in the dermis by the dermis fibroblast, secreted to the outside of the dermis, and polymerized to form collagen fiber after terminal peptide separation under the action of terminal procollagenase. Therefore, the human dermal fibroblast is used as a cell model for researching the promotion of the content of the type I collagen by the polypeptide sodium hyaluronate composite solution, and the effect of promoting the synthesis of the collagen by the polypeptide sodium hyaluronate composite solution is characterized by the up-regulation rate of the type I collagen. The polypeptide sodium hyaluronate complex solutions obtained in examples 1 to 5 and comparative examples 1 to 6 were used to measure the increase rate of type I collagen, and the measurement results are shown in Table 2.

TABLE 2 detection results of elastase inhibition Rate and type I collagen up-regulation Rate

As can be seen from Table 2, the polypeptide sodium hyaluronate composite solution of the present application has the characteristics of higher elastase inhibition rate and type I collagen up-regulation rate, the elastase inhibition rate is 62.8-65.3%, and the type I collagen up-regulation rate is 34.9-36.7%, so that the polypeptide sodium hyaluronate composite solution has the characteristics of good effect of reducing skin wrinkles and strong effect of promoting collagen synthesis, and can play a role in excellent skin tightening and wrinkle resisting effects, increase skin elasticity, and improve skin state.

Comparing example 1 with comparative example 1-3, adding tripeptide-1 into the raw material of the sodium hyaluronate composite solution of the polypeptide of comparative example 1, adding tripeptide-1 derivative into the raw material of the sodium hyaluronate composite solution of comparative example 2, wherein the tripeptide-1 derivative is obtained by treating the tripeptide-1 with ethyl hexadecanoate, adding tripeptide-1 derivative into the raw material of the sodium hyaluronate composite solution of comparative example 3, wherein the tripeptide-1 derivative is obtained by treating the tripeptide-1 with astaxanthin, and adding tripeptide-1 derivative into the raw material of the sodium hyaluronate composite solution of example 1, wherein the tripeptide-1 derivative is obtained by treating the tripeptide-1 with ethyl hexadecanoate and astaxanthin. From this, it can be seen that grafting ethyl palmitate and astaxanthin into tripeptide-1 can effectively increase the elastase inhibition rate and the type I collagen up-regulation.

Comparing example 1 with comparative examples 4-6, the raw material of the comparative example 4 polypeptide sodium hyaluronate complex solution was not added with acetyl hexapeptide-8 and tripeptide-1 derivatives, the raw material of the comparative example 5 polypeptide sodium hyaluronate complex solution was added with acetyl hexapeptide-8, the raw material of the comparative example 6 polypeptide sodium hyaluronate complex solution was added with tripeptide-1 derivatives, and the raw material of the example 1 polypeptide sodium hyaluronate complex solution was added with acetyl hexapeptide-8 and tripeptide-1 derivatives. Therefore, the acetyl hexapeptide-8 and tripeptide-1 derivatives are added into the raw materials at the same time, and the synergistic effect between the acetyl hexapeptide-8 and tripeptide-1 derivatives is utilized, so that the elastase inhibition rate and the type I collagen up-regulation rate can be obviously increased, and the polypeptide sodium hyaluronate composite solution shows better use effect.

(4) The discussion group is composed of 10 professional medical staff, 70 women 35-45 years old are searched for as volunteers, the volunteers sign written informed consent and are randomly divided into 8 groups of 10 persons each, 1 group uses the polypeptide sodium hyaluronate composite solution obtained in example 1, 2-7 groups sequentially uses the polypeptide sodium hyaluronate composite solution obtained in comparative examples 1-6 respectively, the improvement rate of the wrinkle depth, the improvement rate of the wrinkle area, the improvement rate of the elasticity index R2, the improvement rate of the elasticity index R5 and the improvement rate of the elasticity index R7 of the volunteers before and after use are recorded, and the detection results are shown in table 3.

TABLE 3 detection results of anti-wrinkle and skin tightening improvement conditions of skin

As can be seen from table 3, the polypeptide sodium hyaluronate composite solution of the present application had a wrinkle depth improvement ratio of 33.8%, a wrinkle area improvement ratio of 41.3%, an elasticity index R2 improvement ratio of 99.6%, an elasticity index R5 improvement ratio of 98.3%, and an elasticity index R7 improvement ratio of 97.1%. The polypeptide sodium hyaluronate composite solution provided by the application has a better anti-wrinkle and skin tightening effect, and can promote the hydration of skin, enhance the nutrition to dermal cells, improve the plumpness of skin, increase the elasticity and tighten the skin, so that the skin is healthier, better in quality and natural.

Comparing example 1 with comparative examples 1-3, it can be seen that grafting ethyl palmitate, astaxanthin, to tripeptide-1 is beneficial to improving the efficacy of tripeptide-1 derivatives. By combining comparative examples 4 to 6, it can be seen that acetyl hexapeptide-8 and tripeptide-1 derivatives are simultaneously added into raw materials, and the synergistic effect among the acetyl hexapeptide-8 and tripeptide-1 derivatives is utilized to remarkably improve the wrinkle depth, the wrinkle area and the elasticity index and enhance the anti-wrinkle skin tightening effect of the polypeptide sodium hyaluronate composite solution.

It should be noted that the above-described embodiments are only for explaining the present application and do not constitute any limitation of the present application. The application has been described with reference to exemplary embodiments, but it is understood that the words which have been used are words of description and illustration, rather than words of limitation. Modifications may be made to the application as defined in the appended claims, and the application may be modified without departing from the scope and spirit of the application. Although the application is described herein with reference to particular means, materials and embodiments, the application is not intended to be limited to the particulars disclosed herein, as the application extends to all other means and applications which perform the same function.

Claims

1. A polypeptide sodium hyaluronate composite solution for injection, characterized in that: the polypeptide sodium hyaluronate composite solution is composed of a solvent and a solute, the solvent is a buffer solution, and the solute mainly comprises the following raw materials: sodium hyaluronate 5-50 mg/mL, L-carnosine 1-10 mg/mL, acetyl hexapeptide-8 0.05-0.15 mg/mL, acetyl tetrapeptide-5 0.05-0.1 mg/mL, tripeptide-1 derivative 0.03-0.1 mg/mL; the tripeptide-1 derivative is obtained by treating tripeptide-1 with ethyl hexadecanoate and astaxanthin;

The tripeptide-1 derivative is prepared by the following method: an organic solvent and ethyl hexadecanoate are mixed, and then a tripeptide-1 ethanol dilution is added dropwise, and after the addition is complete, the reaction is continued with stirring for 17-23 hours, and then an astaxanthin ethanol dilution is added dropwise, and after the addition is complete, the reaction is continued with stirring for 27-33 hours, and the mixture is concentrated under reduced pressure and dried to obtain a tripeptide-1 derivative;

The weight ratio of hexadecanoic acid ethyl ester, tripeptide-1 in the tripeptide-1 ethanol dilution, and astaxanthin in the astaxanthin ethanol dilution is (4-6): (5-9): (4-6); the dripping time of the tripeptide-1 ethanol dilution is 20-40 min, and the dripping time of the astaxanthin ethanol dilution is 20-40 min;

The total content of the acetyl hexapeptide-8, acetyl tetrapeptide-5 and tripeptide-1 derivatives is greater than 0.15 mg/mL.

2. The polypeptide sodium hyaluronate composite solution for injection according to claim 1, characterized in that the buffer is a phosphate buffer, the pH value of the phosphate buffer is 6.6-7.6, and the osmotic pressure is 270-330 mOsm/L.

3. A polypeptide sodium hyaluronate composite solution for injection according to claim 1, characterized in that the weight ratio of tripeptide-1 to ethanol in the tripeptide-1 ethanol dilution is 1: (3-7), and the weight ratio of astaxanthin to ethanol in the astaxanthin ethanol dilution is 1: (3-7).

4. The polypeptide sodium hyaluronate composite solution for injection according to claim 1, characterized in that the weight ratio of the organic solvent to ethyl hexadecanoate is (30-70): (4-6).

5. The polypeptide sodium hyaluronate composite solution for injection according to claim 1, characterized in that the organic solvent is one or more of ethanol, methanol, propanol and isopropanol.

6. A method for preparing the polypeptide sodium hyaluronate composite solution for injection as claimed in any one of claims 1 to 5, characterized in that it comprises the following steps:

S1. Prepare buffer solution, filter and sterilize to obtain sterile buffer solution;

S2. Under a sterile environment, sterile buffer, sodium hyaluronate, L-carnosine, acetyl hexapeptide-8, acetyl tetrapeptide-5, and tripeptide-1 derivative are mixed to obtain a polypeptide sodium hyaluronate composite solution.